Journal of Biology, Agriculture and Healthcare ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.3, No.

13, 2013

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A Pilot Study on effects of vaccination on immunity of broiler chickens

Ojiezeh, T. I., 1Ophori, E. A., 1Eghafona, N. O., 2Echeonwu, G. O. N., 3Joannis, T. M. and 4Akele, R. Y. 1. Department of Microbiology, University of Benin, Benin City, Edo State, Nigeria. 2. School of Medical Laboratory Science, NVRI, Vom and 3. Department of Regional Laboratory for Avian Influenza and Tran boundary Animal Diseases, NVRI, Vom, Plateau State. 4. Medical Laboratory, Medical Centre, Joseph Ayo Babalola University, Ikeji-Arakeji, Osun State *Correspondence address: pastortksm@yahoo.com, +2348027120080 Abstracts A pilot study was carried out with the aim of highlighting the effects of NDV vaccine on the immune responses of broiler chickens challenged with NDV. Twenty (20) broilers of day-old were used for the study. They were grouped into five of four per group. During the study they were fed with standard feeds and clean water ad libitum. Both vaccinated and unvaccinated groups were challenged with 0.2 saline suspension of 106 ELD50 intradermal inoculation of NDV challenged strain. The vaccinated groups showed neither clinical signs nor symptoms of NDV infections while unvaccinated group showed 100% mortality after 48hr. This result indicate that vaccines is still very important in the prevention, management and control of poultry diseases as maternal immunity passed on to the young chicks at precocial stage could not be relied on to fight against infectious disease in broiler chickens. Therefore, the use of locally produced vaccines should be encouraged among farmers for the prevention, control and management of outbreaks of viral infections in our community. Key: Challenged, Poultry birds- broilers, Newcastle disease virus, Vaccination 1.0 Introduction The problem of viral diseases (pathogenicity) in relation to mortality and morbidity in human population has been a subject of great interest to man. Great scientific strides have been made: the advent of electron microscope has provided us the opportunity to have greater insight to the physiological and anatomical structure of the viruses complex; serology can permit us a peep into molecular changes in an infected host and vaccinology can be relied on to provide a blanket protection against infection by this infective particles. However, the questions of whether we have sufficient knowledge of the disease, its courses and effects so as to develop appropriate tools and interventions for its control and using them to the maximum advantage still remain. The development of the appropriate tools and measures of intervention for the control of diseases falls within the purview of immunomodulation. Vaccination is an effective means to prevent and/or reduce the adverse effects of specific diseases in poultry. Poultry refers to birds that people keep for their use, and generally includes chicken, turkey, duck, goose, quail, pheasant, pigeon, guinea fowl, pea fowl, ostrich, emu and rhea [1]. Disease-causing organisms can be classified, smallest to largest, as viruses, mycoplasma, bacteria, fungi, protozoa, and parasites. All these organisms are susceptible to chemotherapy except viruses. Control of viral diseases is dependent upon prevention through sanitation and biosecurity and by vaccination [1]. Strict sanitation and biosecurity are essential for successful poultry production. Vaccination is no substitute for effective management. It must be understood that vaccines may be effective in reducing clinical disease, but exposed birds, in most cases, still become infected and shed disease organisms [1] The desirable consequences manifest are in form of natural resistance, recovery due to an adaptive immune response and acquired resistance whereas the undesirable consequences are those of autoimmunity, rejection, and hypersensitivity. The disease process sets in as consequences of the failure of the immune system to effectively and efficiently combat invading foreign or non-stop materials that enter the body. The basic function of the immune system is to combat the numerous pathogens present in the body. It is well established that the lowering of the individuals immunity precedes disease conditions [2]. In other words, good health can remain assured as long as the immunity is high. Various substances have been known to have an impact on an individuals immunity, such as drug, nutrition, chemical and immunization [3 - 5].
*1

Journal of Biology, Agriculture and Healthcare ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.3, No.13, 2013

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2.0 Materials and methods 2.1 Source Twenty (20) broiler chickens of day-old from Obasanjo Hatchery, Oluyole Estate, Ibadan, Oyo State were used for this study and transported to Animal house, Achievers University, Owo, Nigeria. 2.2 Innoculums A vial of lyophilized challenged strain of Newcastle disease virus (NDV) obtained from Regional Laboratory for Avian Influenza and Tran boundary Animal Diseases, National Veterinary Research Institute, Vom, Plateau State. The vial was transported under cold chain and standard biosafety practice to Owo, Ondo State. A saline suspension of 106 ELD50 (50 percent Embryo Lethal Dose) were prepared by taken up the vial in 1.5 ml of sterile diluents (physiological saline), then 1 ml of the reconstituted virus was to add 99 ml of sterile normal saline (0.85% NaCl). 2.3 Treatment of broilers They were distributed into five (5) groups of 4 broilers per cage. Groups I, II, III, IV were given intraoccular (eye drops) vaccination within 12 hr. of life using four of patients NDV locally produced vaccines (Biovac for group I, Abic for group II, Hipra for group III and Izovac for group IV) as immune primer. LaSota vaccine (booster dose) was given at day 21 to groups I, II, III and IV. The fifth group was the control chickens, they were not given NDV vaccine either primary or secondary dosages. Meanwhile, all the birds were vaccinated against infectious bursa disease, coccidiosis and administered orally with antibiotics (Doxygen 20/20; Kepro B. V Holland) containing gentamycin sulfate and doxycyline hydrate at the dose rate of 1g per 3L of drinking water for the first five days). The broilers were housed in battery cages of 0.301 m2 / bird as recommended by Mustafa et al. [6] in the experimental animal house of Biological Science Department, College of Natural and Applied Sciences, Achievers University, Owo. All experimental protocols complied with NIH guidelines [7], as approved by the ethical and research committee, Achievers University, Owo. All the groups were challenged intramuscularly with 0.2 ml saline suspension of 106 ELD50 at five weeks old. Their physiological response to the viral inoculums was observed; the stool frequency (diarrhea), difficult breathing and nasal discharge, and weight changes were specifically noted. During the entire study the broilers were fed with feeds compounded carefully to meet 23% crude protein (CP) and 3200K.cal. metabolizable energy (ME) for broiler starter and 20% CP and 3000K.cal. for broiler finisher. We ensured that the levels of mycotoxins in feed were maintained relatively low throughout the experiment, the quality and quantities of groundnut cake (GNC), soya bean and rice bran included in the feeds were the same for both starter and finisher mash. The percentage CP in feed and ingredients were determined by the burette method [8], while the metabolizable energy was determined by the Bomb calorimeter method [9]. 3.0 Result and Discussion The study shows that all the broilers of control group were knock-dead by the challenge within 48 hours, while all the NDV vaccinated groups survived the challenge. There were no paralysis and other clinical signs and symptoms of NDV infection. There were no observable changes in the physical activities of the vaccinated groups which indicate that, the four locally produced vaccines from NVRI, Vom that were used in this study were potent vaccines against the challenged strain. Those vaccines may have conferred on the tested bird in groups I - IV, the ability to resist the non self. This finding is in tandem with the report of Banu et al. [10] and Chowhdury et al. [11] which supported the use of live vaccine to enhance immunity. The surviving birds further highlight the significances of vaccination especially in poultry management. Live vaccines enhanced their immune responses; the administration of secondary dosage to the chicks following primer dosage may have enhanced their immunity. The top up dosage would have prompted secondary responses in which there would be a rapid response by memory B-lymphocyte cell resulting in marked increase in antibody production (titre) in the birds. This principle is often used in active immunization against infectious diseases in poultry management. Little wonder what happens to maternal antibody pass - on to the chicks at precocial stage, it should have had protective effect on the defenseless young species at least for a time. According to King et al. [12] immunemediated maternal effects are believed to play an integral role in the disease resistance of mammalian [1314] and avian offspring [1518]. Maternal antibodies passively immunize immunologically nave young against virulent antigens and parasites that the offspring might encounter in its immediate developmental environment [19-20], yet the effects of maternal antibodies on offspring development are not well defined for these altricial-developing species [21]. Maternal antibodies are transferred across the follicular epithelium into the yolk during oogenesis [21-23]. There are three classes of avian immunoglobulins (IgY, IgM and IgA). Of these, IgY is transferred at the highest concentration and is functionally homologous to mammalian IgG [24]. IgA and IgM are found predominantly in the egg white of chicken eggs, but have been detected in the yolk at low concentration [25-26]. Prior to hatch, maternal IgY is absorbed into embryonic circulation [23], where it confers passive immunity to immunologically immature hatchlings [16, 17, 20, 24, 27]. IgM is also absorbed into circulation, though at low

Journal of Biology, Agriculture and Healthcare ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.3, No.13, 2013

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concentrations (<1%) [24]. There is therefore, the need to study further the maternal antibody to determine each specific antibody concentration and it half life in broiler chickens.

Table 1: Clinical signs and Physiological changes of vaccinated and unvaccinated broiler chickens following NDV challenge and control group Treatment groups I II III IV Control Weight (kg) ) Diarrhea b/4 challenge Difficult breathing/ Nasal discharge +++ Physical activities Paralysis (%) Mortality (%) Weight difference(kg) (48h after challenge) 0 0 0.020 0 0 0.030 0 0 0.021 0 0 0.019 100 100 -0.040

0.710 0.665 0.570 0.606 0.636

+++

100

Conclusion This study further highlights the relevance of vaccination in disease prevention, control and management. Vaccines produce locally are potent against NDV challenged strain. And the use of vaccines should be encouraged among farmers for control and management of outbreak of Newcastle disease Virus in our locality. Conflict of Interests The authors do not have a direct financial relationship with the commercial identity mentioned in this paper. Acknowledgments The authors wish to acknowledge Animal Scientist; Mr. Ademola Obasemola, Animal House Unit and Dr. P. A. Okiki of Microbiology Department, Achievers University, Owo, Ondo State for their technical support. References 1. Jacob, J. P., Butcher, G. D. and Mather, F. B. (1998). The document no PS 36. Animal Science Department, Florida cooperative Extension Service, Institute of Food and Agriculture Science. University of Florida. Reviewed 2012. 2. Ojiezeh, T. I., Ibeh, N. I. and Okoko, F. J. (2011). Effects of cations on the immune indices of Salmonella enterica challenged rabbits. Tanzania Journal of Natural and Applied Sciences 2 (1):304 309. Chevaliar, P., Sevilla, R., Zakes, L., Segas, E., Belmonte, G., Parent, G. and Jambou, B. (1996). Immunonutritional recovery of children with severe malnutrition. Sante 64: 201 - 8. Rahman, M. J., Sarkar, P., Roy, K. S., Ahmed, M. S., Christi, J., Azim, T., Mattan, M., Sack, D., Anderson, J. and Ragib, R. (2005). Effects of zinc supplementation as adjunct therapy on the systemic immune responses in Shigellosis. American Journal of Clinical Nutrition 81 (2): 495-502. Turnlund, H. R., Jacob, R. A., Keen, C. L., Strain, J. J., Kelly, D. S., Domek, J. M., Keyes, W. R., Ensunsa, J. L., Lykkesfeidt, J. and Coulter, J. (2004). Longterm high copper intake effects on indexes of copper status, antioxidant status and immune function in young men. American Journal of Clinical Nutrition 79:1037-44.

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6. Mustafa, Y. M., Muhammed, A. M., Aflab, A. A. and Mansoir-ud-Din-Ahmed. (2010). Influence of stocking Density on immune response of broiler against Newcastle Disease Virus. Pakistan Journal of Life and Social Sciences 8(1):7-10. 7. National Research Council (1985) Guides for the Care and Use of Laboratory Animals, National Institutes of Health, Bethesda, Md, USA. 8. Ranjna, C. (1999). Practical Clinical Biochemistry Method and Interpretation, Jaypee Brother Medical publishers Ltd. New Delhi, India. 267pp. 9. AOAC. (1980). Official methods of analysis of the association of official analytical chemists.11th Edition Washington D.C, U.S.A. 595pp.

Journal of Biology, Agriculture and Healthcare ISSN 2224-3208 (Paper) ISSN 2225-093X (Online) Vol.3, No.13, 2013

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Banu, N. A., Islam, M. S., Chowdhury, M. M. H. and Islam, M. A. (2009). Determination of immune response of Newcastle disease virus vaccines in layer chickens. Journal of the Bangladesh Agricultural University 7(2): 329 - 334. 11. Chowdhury, T. I. M. F. R., Sarker, A. J., Amin, M. M. and Hossain, W. I. M. A. (1981). Studies on Newcastle disease in Bangladesh, Pp 12 - 20. In: Gibba, E. P. J. (ed). Virus. Disease of Food Animal Vol. 1: A Research Report, Sec 2. The role of residual maternal antibody on immune response and selection of an optimal age for primary vaccination of chicks. 12. King, M.O., Owen, J.P. and Schwabl, H. G. (2010). Are Maternal Antibodies Really That Important? Patterns in the Immunologic Development of Altricial Passerine House Sparrows (Passer domesticus). PLoS ONE 5(3): e9639. doi:10.1371/journal. pone. 0009639 13. Lyche, J.L., Larsen, H.J.S., Skaare, J.U., Tverdal, A., Johansen, G.M. and Ropstad, E., (2006). Perinatal exposure to low doses of PCB 153 and PCB 126 affects maternal and neonatal immunity in goat kids. Journal of Toxicology and Environmental Health-Part a-Current Issues 69: 139-158. 14. Anderson, R. W. (1995). On the maternal transmission of immunity - a molecular attention hypothesis. Biosystems 34: 87105. 15. Chaffer, M., Schwartsburd, B. and Heller, E. D. (1997). Vaccination of turkey poults against pathogenic Escherichia coli. Avian Pathol. 26: 377390. 16. Fahey, K.J., Crooks, J.K. and Fraser, R.A. (1987). Assessment by ELISA of passively acquired protection against infectious bursal disease virus in chickens. Aust. Vet. J. 64: 203207. 17. Wang, Y.W., Sunwoo, H., Cherian, G. and Sim, J. S. (2004). Maternal dietary ratio of linoleic acid to alphalinolenic acid affects the passive immunity of hatching chicks. Poult. Sci. 83: 20392043. 18 Grindstaff, J.L., Hasselquist, D., Nilsson, J.A., Sandell, M., Smith, H. G. and Stjernman, M. (2006). Transgenerational priming of immunity: maternal exposure to a bacterial antigen enhances offspring humoral immunity. Proceedings of the Royal Society B-Biological Sciences 273: 25512557. 29. Gasparini, J., McCoy, K. D., Staszewski, V., Haussy, C. and Boulinier, T. (2006). Dynamics of anti-Borrelia antibodies in Blacklegged Kittiwake (Rissa tridactyla) chicks suggest a maternal educational effect. Canadian Journal of Zoology-Revue Canadienne De Zoologie 84: 623627. 20. Grindstaff, J. L. (2008). Maternal antibodies reduce costs of an immune response during development. J. Exp. Biol. 211: 654660. 21. Apanius, V. (1998). Ontogeny of immune function. pp. 203222. In: Stark, J. M., Ricklefs, R. E., editors. Avian Growth and Development Oxford: Oxford University Press. 22. Loeken, M. R. and Roth, T. F. (1983). Analysis of maternal IgG subpopulations which are transported into the chicken oocyte. Immunology 49: 2128. 23. Kowalczyk, K., Daiss, J., Halpern, J. and Roth, T. F. (1985). Quantitation of maternal-fetal IgG transport in the chicken. Immunology 54: 755762. 24. Hamal, K. R., Burgess, S.C., Pevzner, I. Y. and Erf, G. F. (2006). Maternal antibody transfer from dams to their egg yolks, egg whites, and chicks in meat lines of chickens. Poult. Sci. 85: 13641372. 25. Kaspers, B., Schranner, I. and Losch, U. (1991). Distribution of immunoglobulins during embryogenesis in the chicken. Journal of Veterinary Medicine Series a-Zentralblatt Fur Veterinarmedizin Reihe a-Physiology Pathology Clinical Medicine 38: 7379. 26.Yamamoto, H., Watanabe, H., Sato, G., Mikami, T. (1975). Identification of immunoglobulins in chicken eggs and their antibody activity. Jpn. J. Vet. Res. 23: 131140. 27. Heller, E. D., Leitner, G., Drabkin, N. and Melamed, D. (1990). Passive-immunization of chicks against Escherichia-coli. Avian Pathol. 19: 345354.

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